The present invention relates to a method for breeding an organism and a feed.
Recently, culture of useful organisms such as domestic animals, laboratory animals, domestic fowls, fishes and shrimps have remarkably become popular. On the other hand, occurrence of various diseases, underdevelopment and the like caused by stress due to a breeding environment such as packed breeding have become problems.
The main object of the present invention is to provide a method for breeding an organism and a feed useful for preventing or treating diseases of useful organisms such as cultured animals and for maintaining the health of the useful organisms.
The present invention is outlined as follows. The first invention of the present invention relates to a method for breeding an organism, characterized in that the method comprises administering:
(1) a heat treatment product of at least one compound selected from the group consisting of (a) uronic acid or a uronic acid derivative; (b) a saccharide containing uronic acid and/or a uronic acid derivative; and (c) a material containing a saccharide containing uronic acid and/or a uronic acid derivative;
(2) 4,5-dihydroxy-2-cyclopenten-1-one of formula (I): 
xe2x80x83and/or
(3) a derivative of 4,5-dihydroxy-2-cyclopenten-1-one of formula (I), to an organism.
The second invention of the present invention relates to a feed for an organism which contains, which is produced by adding thereto, and/or which is produced by diluting a heat treatment product of at least one compound selected from the group consisting of (a), (b) and (c) as described above; 4,5-dihydroxy-2-cyclopenten-1-one of formula (I); and/or a derivative of 4,5-dihydroxy-2-cyclopenten-1-one of formula (I).
The third invention of the present invention relates to a composition for breeding an organism which contains a heat treatment product of at least one compound selected from the group consisting of (a), (b) and (c) as described above; 4,5-dihydroxy-2-cyclopenten-1-one of formula (I); and/or a derivative of 4,5-dihydroxy-2-cyclopenten-1-one of formula (I).
In the first to third inventions of the present invention, the organism is, for example, a cultured animal or a pet animal. The cultured organism is exemplified by a domestic animal, a laboratory animal, a domestic fowl, a fish, a crustacean and a shellfish. The heat treatment product is exemplified by a material containing 4,5-dihydroxy-2-cyclopenten-1-one of formula (I).
The feed of the second invention of the present invention is exemplified by a feed for improving physical condition.
The composition for breeding an organism of the third invention of the present invention is exemplified by a composition for soaking an organism, an additive for a feed, and an additive for a drink.
As used herein, a heat treatment product of at least one compound selected from the group consisting of uronic acid or a uronic acid derivatives a saccharide containing uronic acid and/or a uronic acid derivative; and a material containing a saccharide containing uronic acid and/or a uronic acid derivative is not limited to a specific one as long as it has an effect of increasing efficiency of breeding an organism such as viability, fattening rate, spawning rate, birthrate or weaning rate.
Uronic acid is (also called as glycuronic acid) a generic name for hydroxyaldehyde-acids in which the aldehyde group of aldose is kept intact and only the primary alcohol group at the other end is oxidized into a carboxyl group. Uronic acid naturally exists as constituents of various polysaccharides in animals and plants.
Examples of uronic acids which can be used in the present invention include, but are not limited to, galacturonic acid, glucuronic acid, gluronic acid, mannuronic acid and iduronic acid. Derivatives of uronic acid include lactones, esters, amides and salts thereof.
As used herein, examples of saccharides containing uronic acid and/or a uronic acid derivative include, but are not limited to, pectin, pectic acid, alginic acid, hyaluronic acid, heparin, heparan sulfate, fucoidan, chondroitin sulfate, chondroitin and dermatan sulfate. Decomposition products (products of chemical, enzymatic or physical treatment) of the above-mentioned saccharides as well as derivatives and salts of the decomposition products can also be used.
As used herein, a material containing a saccharide containing uronic acid and/or a uronic acid derivative is not limited to a specific one as long as the material contains the above-mentioned saccharide containing uronic acid and/or a uronic acid derivative. Materials containing a saccharide containing uronic acid and/or a uronic acid derivative derived from plants, algae, microorganisms and animals can be used.
Alternatively, a processed agricultural or marine food product containing uronic acid and/or a uronic acid derivative may be used directly or after drying and grinding as a raw material for the heat treatment product in the present invention.
A method used for the heat treatment in the production of the heat treatment product of the present invention is not limited to a specific one as long as it produces a heat treatment product having physiological activities such as an activity of increasing breeding efficiency and an activity of improving physical condition. At least one compound selected from the group consisting of uronic acid or a uronic acid derivative; a saccharide containing uronic acid and/or a uronic acid derivative; and a material containing a saccharide containing uronic acid and/or a uronic acid derivative may be heated, for example, at 60-350xc2x0 C. for a few seconds to a few days, preferably at 80-150xc2x0 C. for a few minutes to a few days. A heat treatment product used in the present invention can be obtained by heating pectin or alginic acid, for example, at 80-150xc2x0 C. for a few minutes to a few days. Furthermore, a heat treatment product of interest can be obtained by heating uronic acid, a lactone of uronic acid or a uronic acid ester at 60-150xc2x0 C. for a few minutes to a few days.
Either wet heating or dry heating may be used for the heat treatment in the present invention. Among these, wet heating is preferable in view of production efficiency of a compound contained in the heat treatment product of the present invention having physiological activities such as an activity of increasing breeding efficiency and an activity of improving physical condition, for example, 4,5-dihydroxy-2-cyclopenten-1-one of formula (I) (hereinafter simply referred to as cyclopentenone). Any wet heating such as steam heating, pressurized steam heating and pressurized heating can be used. Direct heating using dry hot wind and indirect heating through a division wall from a heat source can be used as dry heating. Direct heating is exemplified by air current dry heating and spray dry heating. Drum dry heating and the like can be used as indirect heating.
Any methods may be used for producing cyclopentenone of the present invention. Cyclopentenone may be synthesized according to a chemical synthesis method [Carbohydrate Res., 247:217-222 (1993); Helvetica Chimica Acta, 55:2838-2844 (1972)]. Cyclopentenone produced in a heat treatment product of at least one compound selected from the group consisting of uronic acid or a uronic acid derivative; a saccharide containing uronic acid and/or a uronic acid derivative; and a material containing a saccharide containing uronic acid and/or a uronic acid derivative, or a product purified therefrom can be used. These heat treatment products containing cyclopentenone as well as products partially purified or purified therefrom can be used in the present invention (WO 98/13328).
A method used for the heat treatment in the production of the heat treatment product containing cyclopentenone of the present invention is not limited to a specific one as long as it produces cyclopentenone. Uronic acid or a uronic acid derivative; a saccharide containing uronic acid and/or a uronic acid derivative; or a material containing a saccharide containing uronic acid and/or a uronic acid derivative may be heated, for example, at 60-350xc2x0 C. for a few seconds to a few days, preferably at 80-150xc2x0 C. for a few minutes to a few days. A heat treatment product containing cyclopentenone can be obtained by heating pectin or alginic acid, for example, at 80-150xc2x0 C. for a few minutes to a few days. Furthermore, a heat treatment product containing cyclopentenone of interest can be obtained by heating uronic acid, a lactone of uronic acid or a uronic acid ester at 60-150xc2x0 C. for a few minutes to a few days.
Although the pH used for the heat treatment is not limited to a specific one, it is preferable to carry out the heat treatment under neutral to acidic conditions. The pH for the heat treatment may be adjusted depending on the raw material used. The production of cyclopentenone is usually accelerated by heating under acidic conditions.
The concentration of the raw material during the heat treatment is within any range in which cyclopentenone can be produced by heat treatment. The concentration may be determined in consideration of operationality, yield and the like.
Either wet heating or dry heating may be used for the heat treatment in the present invention. Among these, wet heating is preferable in view of production efficiency of cyclopentenone. Any wet heating such as steam heating, pressurized steam heating and pressurized heating can be used. Direct heating using dry hot wind and indirect heating through a division wall from a heat source can be used as dry heating. Direct heating is exemplified by air current dry heating and spray dry heating. Drum dry heating and the like can be used as indirect heating.
Cyclopentenone in the heat treatment product used in the present invention may be collected by known purification/isolation means including chemical means and physical means. Conventional purification means such as gel filtration, fractionation using a molecular weight fractionating membrane, solvent extraction, fractional distillation and various chromatographies (ion-exchange resins, normal phase or reverse phase) can be used in combination to collect cyclopentenone produced in the heat treatment product.
For example, cyclopentenone is produced in a heat treatment product by heating a 1% solution of D-glucuronic acid as uronic acid at 121xc2x0 C. for 4 hours. Cyclopentenone in the heat treatment product is extracted with a solvent. The extract is concentrated. The concentrate is then separated on silica gel column chromatography. Eluted fractions containing cyclopentenone are concentrated. Cyclopentenone is extracted from the concentrate with chloroform, thereby isolating cyclopentenone in the heat treatment product.
Furthermore, cyclopentenone is purified by subjecting the heat treatment product of glucuronic acid to an ion-exchange resin column, preferably an anion-exchange resin column and collecting non-adsorptive fractions. Alternatively, purified cyclopentenone can be obtained by the following steps. The heat treatment product of glucuronic acid is subjected to an activated carbon column. Non-adsorptive fractions are removed. The column is washed. Elution is carried out using a hydrophilic organic solvent (e.g., an aqueous ethanol solution, preferably an aqueous ethanol solution at a concentration of 40% or more). In addition, highly purified cyclopentenone can be obtained by combining these methods.
A derivative of 4,5-dihydroxy-2-cyclopenten-1-one (hereinafter simply referred to as a cyclopentenone derivative) can also be used in the present invention. The cyclopentenone derivative is not limited to a specific one as long as the objects of the present invention can be accomplished by using the derivative. Cyclopentenone derivatives of formulas (II) to (V) are exemplified: 
wherein the bond in the five-membered ring represented by a broken line means that the five-membered ring may be either a cyclopentene ring having a double bond or a saturated cyclopentane ring; if the five-membered ring is a cyclopentene ring, X1 is OH, Y1 is xe2x95x90O and Z1 is H; if the five-membered ring is a cyclopentane ring, X1 is xe2x95x90O, Y1 is OH and Z1 is OH; W1 is a residue in which an SH group is removed from an SH group-containing compound; 
wherein R1 and R2 may be identical or different each other, and are hydrogen, an aliphatic group, an aromatic group or an aromatic aliphatic group; 
wherein R3 and R4 may be identical or different each other, and are hydrogen, an aliphatic group, an aromatic group or an aromatic aliphatic group, provided that R3 and R4 are not simultaneously H; and 
wherein the bond in the five-membered ring represented by a broken line means that the five-membered ring may be either a cyclopentene ring having a double bond or a saturated cyclopentane ring; if the five-membered ring is a cyclopentene ring, X2 is OR5, Y2 is xe2x95x90O and Z2 is H; if the five-membered ring is a cyclopentane ring, X2 is xe2x95x90O, Y2 is OR6 and Z2 is OR7; R5 is R8 or xe2x80x94(CO)xe2x80x94R9; R6 is H, R10 or xe2x80x94(CO)xe2x80x94R11; R8 is H, R12 or xe2x80x94(CO)xe2x80x94R13 (R8, R9, R10, R11, R12 and R13 may be identical or different each other, and are an aliphatic group, an aromatic group or an aromatic aliphatic group; R9 , R11 and R13 may be H), provided R6 and R7 are not simultaneously H; W2 is a residue in which an SH group is removed from an SH group-containing compound.
Optical isomers of the materials containing the cyclopentenone derivatives or salts thereof can be used in the present invention.
The cyclopentenone derivative of formula (II) is described in detail in WO 98/39291. The derivative can be obtained by reacting cyclopentenone with an SH group-containing compound such as cysteine or glutathione. A material containing the cyclopentenone derivative can be obtained by adding an SH group-containing compound to a material containing cyclopentenone. The derivatives are exemplified by compounds of formulas (VI) to (IX): 
The compound of formula (VI) is a product of reaction between cyclopentenone and glutathione under acidic conditions. The compound of formula (VII) is a product of reaction between cyclopentenone and glutathione under neutral conditions. The compound of formula (VIII) is a product of reaction between cyclopentenone and cysteine under acidic conditions. The compound of formula (IX) is a product of reaction between cyclopentenone and cysteine under neutral conditions.
The cyclopentenone derivative of formula (III) is described in detail in WO 98/40346 and Japanese Patent Application No. 10-231659. The derivative can be obtained by simultaneously or sequentially reacting cyclopentenone with a carboxylic acid having an aliphatic group, an aromatic group or an aromatic aliphatic group and/or a reactive derivative thereof. The cyclopentenone derivative is exemplified by diacetylcyclopentenone, dibenzoylcyclopentenone, dihexanoylcyclopentenone, dimyristoylcyclopentenone, dioctanoylcyclopentenone, di-3-octenoylcyclopentenone, dibutyrylcyclopentenone, didecanoylcyclopentenone, divalerylcyclopentenone, dipropionylcyclopentenone, di-2-hexenoylcyclopentenone, diisobutyrylcyclopentenone, dimethoxyacetylcyclopentenone, methoxyfumarylcyclopentenone and methoxymaleylcyclopentenone. Formula (X) below represents the structure of dipropionylcyclopentenone. Formula (XI) below represents the structure of dibenzoylcyclopentenone. 
The cyclopentenone derivative of formula (IV) is described in detail in WO 99/00349. The derivative can be obtained by simultaneously or sequentially reacting cyclopentenone with an alcohol having an aliphatic group, an aromatic group or an aromatic aliphatic group and/or a reactive derivative thereof. The cyclopentenone derivative is exemplified by 4-benzylcyclopentenone ether, 5-benzylcyclopentenone ether, 4,5-dibenzylcyclopentenone ether, 4-t-butylcyclopentenone ether, 5-t-butylcyclopentenone ether and 4,5-di-t-butylcyclopentenone ether. Formulas (XII) and (XIII) below represent the structures of 5-benzylcyclopentenone ether and 4,5-di-t-butylcyclopentenone ether, respectively. 
The cyclopentenone derivative of formula (V) is described in detail in Japanese Patent Application No. 10-232746. The derivative can be obtained by reacting the compound of formula (III) or the compound of formula (IV) with an SH group-containing compound such as cysteine or glutathione. Examples of the derivatives include compounds of formulas (XIV) and (XV) below. 
The compound of formula (XIV) is a product of reaction between dihexanoylcyclopentenone and glutathione. The compound of formula (XV) is a product of reaction between 4,5-di-t-butylcyclopentenone ether and glutathione.
A cyclopentenone derivative having an xcex1,xcex2 unsaturated carbonyl structure can be preferably used in the present invention.
The heat treatment product, cyclopentenone or a cyclopentenone derivative used in the present invention has physiological activities such as a carcinostatic activity, an antiproliferation activity against tumor cells, an apoptosis-inducing activity, an activity of inhibiting topoisomerase II, an activity of inducing differentiation in tumor cells, an antirheumatic activity, an activity of inhibiting rheumatoid arthritis, an activity of inducing Fas antigen production, an antimicrobial activity, an antiviral activity, an activity of improving liver function, an activity of inducing heat shock protein, an activity of normalizing blood component, an activity of enhancing tumor immunity, an anti-inflammatory activity, an activity of inhibiting tumor necrosis factor production, an activity of inhibiting nitrogen monoxide production and immunoregulatory activities such as an activity of inhibiting delayed hypersensitivity, an activity of inhibiting lymphocyte blastogenesis, an activity of inhibiting mixed lymphocyte reaction, an activity of inhibiting IgE production and an activity of inhibiting carrageenan edema. Based on these activities, the feed of the present invention which contains the heat treatment product, cyclopentenone and/or a cyclopentenone derivative used in the present invention as an active ingredient is very useful as a feed for improving physical condition of an organism such as a feed for activating biological defense mechanism, a feed for activating antibody production mechanism, a feed for normalizing saccharide metabolism or a feed effective in protecting and preventing infection with pathological organism.
The cyclopentenone compound of formula (II) produced from cyclopentenone and an SH group-containing compound is produced as a metabolite in an organism in the presence of cyclopentenone and an SH group-containing compound. The feeds of the present invention include a feed that contains cyclopentenone and an SH group-containing compound such as cysteine or glutathione and in which the cyclopentenone derivative is produced in the feed or in a living body. Such a feed is very effective as the feed of the present invention.
The daily dosage of the heat treatment product of at least one compound selected from the group consisting of uronic acid or a uronic acid derivative; a saccharide containing uronic acid and/or a uronic acid derivative; and a material containing a saccharide containing uronic acid and/or a uronic acid derivative (hereinafter simply referred to as the heat treatment product of the present invention) is usually 1 to 20000 mg per kg body weight of the organism of interest. The heat treatment product can be added to and mixed with raw materials for an artificial mixed feed. Alternatively, it can be mixed with powder raw materials for an artificial mixed feed, and then added to and mixed with other raw materials.
A daily dosage of cyclopentenone and/or a derivative thereof is usually 0.01 to 200 mg per kg body weight of the organism of interest (e.g., a cultured animal). It can be added to and mixed with raw materials for an artificial mixed feed. Alternatively, it can be mixed with raw materials of powder form for an artificial mixed feed, and then added to and mixed with other raw materials. At least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention can be directly added to and mixed with a feed for the organism of interest.
The content of the heat treatment product of the preset invention in the feed for the organism of interest is not limited to a specific one and may be determined depending on the objects. The suitable content is 0.05 to 50 w/w %.
The content of cyclopentenone and/or a derivative thereof in the feed for the organism of interest is not limited to a specific one and may be determined depending on the objects. The suitable content is 0.001 to 1 w/w %.
Artificial mixed feeds include those produced using raw materials derived from animal such as fish powder, casein and squid meal, raw materials derived from plant such as soybean cake, wheat flour, starch and feed yeast, animal oil such as cod-liver oil and squid-liver oil, vegetable oil such as soybean oil and rape-seed oil, vitamins, minerals, amino acids, antioxidants and the like as raw materials. The artificial mixed feeds also include feeds for fishes such as minced fish meat.
The method for producing the feed of the present invention is not limited to a specific one as long as the produced feed contains, is produced by adding thereto and/or is produced by diluting at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention.
At least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention can be administered to an organism of interest by directly adding the compound to water or seawater in a pool, a water tank, a reservoir tank or a breeding area and soaking the organism therein. This soaking method is particularly effective when the intake of feed by an organism of interest is reduced.
The concentration of cyclopentenone and/or a derivative thereof in water or seawater is not limited to a specific one and may be determined depending on the objects. The suitable concentration is 0.00001 to 0.01 w/w %.
The concentration of the heat treatment product of the present invention in water or seawater is not limited to a specific one and may be determined depending on the objects. The suitable concentration is 0.005 to 5 w/w %.
Furthermore, a drink containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention may be given to an organism of interest as a drink for breeding.
The concentration of cyclopentenone and/or a derivative thereof in a drink is not limited to a specific one and may be determined depending on the objects. The suitable concentration is 0.0001 to 1 w/w %.
The concentration of the heat treatment product of the present invention in a drink is not limited to a specific one and may be determined depending on the objects. The suitable concentration is 0.005 to 5 w/w %.
A composition for breeding an organism containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention as an active ingredient (e.g., a composition for soaking an organism, an additive for a feed and an additive for a drink) may be produced according to conventional methods.
Organisms to which the present invention can be applied include, but are not limited to, cultured animals such as domestic animals (e.g., horses, cows, pigs, sheep, goats, camels and llamas), laboratory animals (e.g., mice, rats, guinea pigs and rabbits), domestic fowls (e.g., chickens, ducks, turkeys and ostriches), fishes (e.g., red sea breams, parrot fishes, bastard halibuts, flatfishes, yellowtails, young yellowtails, amberjacks, tunas, yellow jacks, sweetfishes, salmon, trout, tiger globefishes, eels, lochs and catfishes), crustaceans (e.g., prawns, black tiger shrimps, yellow sea prawns and blue crabs) and shellfishes (e.g., abalones, turban shells, scallops and oysters) as well as pet animals (e.g., dogs and cats). The present invention can be widely applied to land animals and aquatic animals.
No death was observed when cyclopentenone or a derivative thereof was orally administered to a mouse at a single dosage of 100 mg/kg. Furthermore, no death was observed when the heat treatment product of the present invention was orally administered to a mouse at a single dosage of 300 mg/kg.
Bacterial infection and viral infection in an organism of interest such as a domestic animal, a laboratory animal, a domestic fowl, a fish, a crustacean, a shellfish or a pet animal is prevented or treated. Such prevention or treatment can be accomplished by giving a feed containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention to the organism. Alternatively, the prevention or the treatment can be accomplished by soaking the organism in a solution containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention. Furthermore, disease states in an infected organism is remarkably ameliorated. Additionally, the health of an organism of interest is maintained, resulting in remarkable improvement in viability, fattening rate, spawning rate, birthrate, weaning rate, growth rate or the like.
Cultured animals had the following problems. (1) Diseases due to bacterial infection frequently occur. Since the organisms are cultured in a limited area, all of them will be infected and die in a short period of time once an infectious disease occurs. (2) The organisms are susceptible to parasite infections, nutritional diseases, environmental diseases and tumors. (3) Great stress due to the narrow breeding area causes the cultured organisms to rub the body surface to the breeding facilities and be scratched. The scratches make bacteria and parasites easy to adhere to the animals. (4) Intake of feed decreases due to the stress, resulting in retarded growth. The feed for improving physical condition of the present invention can greatly decrease the stress in the cultured animal bred in a narrow area, prevent the rubbing of the body surface to breeding facilities, make the appetite of the organism better, and remarkably increase growth rate, birthrate, spawning rate, weaning rate, disease prevention rate and the like based on its various physiological activities such as an antimicrobial activity, an anti-inflammatory activity and an antioxidant activity.
The present invention provides a feed for a land animal or an aquatic animal and a solution for soaking an organism containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention. The feed and the solution are very useful as an antibacterial feed, an antiviral feed, an antibacterial solution for soaking an organism and an antiviral solution for soaking an organism.
The present invention provides a method for breeding a land animal or an aquatic animal, and a method for treating or preventing a disease in a land animal or an aquatic animal such as an infectious disease caused by a pathological microorganism such as a bacterium, a fungus or a virus, characterized in that the method comprises administering at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention to the animal.
Furthermore, the present invention provides a method for breeding an organism, or treating or preventing a disease, characterized in that the method comprises soaking a land animal or an aquatic animal in an aqueous solution containing at least one compound selected from the group consisting of cyclopentenone, a derivative thereof and the heat treatment product of the present invention.
The method of the present invention is remarkably effective in treating or preventing infections diseases in cultured fishes (e.g., infection of bastard halibuts with lymphocystis virus, infection with gliding bacteria, and white-mouth disease, an infectious disease in globefishes). When onset or sign of these infectious diseases are recognized, loss of commercial value or occurrence of damage due to these diseases can be prevented by using the feed of the present invention or the method for soaking an organism of the present invention. Furthermore, conventionally used antibiotics and formalin are not required according to the present invention. Thus, problems associated with the residual toxicity thereof are solved.